Pro-fluorescent mitochondria-targeted real-time responsive redox probes synthesised from carboxy isoindoline nitroxides: Sensitive probes of mitochondrial redox status in cells.
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Authors
Chong, Kok Leong
Cullen, Jason K
Morrow, Benjamin J
Fairfull-Smith, Kathryn E
New, Elizabeth J
Murphy, Mike
Publication Date
2018-11Journal Title
Free radical biology & medicine
ISSN
0891-5849
Publisher
Elsevier
Volume
128
Pages
97-110
Language
eng
Type
Article
This Version
AM
Physical Medium
Print-Electronic
Metadata
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Chong, K. L., Chalmers, B. A., Cullen, J. K., Kaur, A., Kolanowski, J. L., Morrow, B. J., Fairfull-Smith, K. E., et al. (2018). Pro-fluorescent mitochondria-targeted real-time responsive redox probes synthesised from carboxy isoindoline nitroxides: Sensitive probes of mitochondrial redox status in cells.. Free radical biology & medicine, 128 97-110. https://doi.org/10.1016/j.freeradbiomed.2018.03.008
Abstract
Here we describe new fluorescent probes based on fluorescein and rhodamine that provide reversible, real-time insight into cellular redox status. The new probes incorporate bio-imaging relevant fluorophores derived from fluorescein and rhodamine linked with stable nitroxide radicals such that they cannot be cleaved, either spontaneously or enzymatically by cellular processes. Overall fluorescence emission is determined by reversible reduction and oxidation, hence the steady state emission intensity reflects the balance between redox potentials of critical redox couples within the cell. The permanent positive charge on the rhodamine-based probes leads to their rapid localisation within mitochondria in cells. Reduction and oxidation also leads to marked changes in the fluorophore lifetime, enabling monitoring by fluorescence lifetime imaging microscopy. Finally, we demonstrate that administration of a methyl ester version of the rhodamine-based probe can be used at concentrations as low as 5 nM to generate a readily detected response to redox stress within cells as analysed by flow cytometry.
Keywords
Cells, Cultured, Mitochondria, Fibroblasts, Humans, Colorectal Neoplasms, Nitrogen Oxides, Antioxidants, Fluorescent Dyes, Microscopy, Fluorescence, Oxidation-Reduction, Molecular Imaging
Sponsorship
MRC (MC_UU_00015/3)
Wellcome Trust (110159/Z/15/Z)
Identifiers
External DOI: https://doi.org/10.1016/j.freeradbiomed.2018.03.008
This record's URL: https://www.repository.cam.ac.uk/handle/1810/277179
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