Mitochondrial membrane potential (ΔΨm) is a universal selective indicator of mitochondrial function and is known to play a central role in many human pathologies, such as diabetes mellitus, cancer and Alzheimer’s and Parkinson’s diseases. Here, we report the design, synthesis and several applications of mitochondria-activatable luciferin (MAL), a bioluminescent probe sensitive to ΔΨm, and partially to plasma membrane potential (ΔΨp), for non-invasive, longitudinal monitoring of ΔΨm in vitro and in vivo. We applied this new technology to evaluate the aging-related change of ΔΨm in mice and showed that nicotinamide riboside (NR) reverts aging-related mitochondrial depolarization, revealing another important aspect of the mechanism of action of this potent biomolecule. In addition, we demonstrated application of the MAL probe for studies of brown adipose tissue (BAT) activation and non-invasive in vivo assessment of ΔΨm in animal cancer models, opening exciting opportunities for understanding the underlying mechanisms and for discovery of effective treatments for many human pathologies.
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The data that support the findings of this study are available from the corresponding author upon request. CCDC nos. 1940412, 1940411, 1940410 and 1940409 for compounds 7, 9, 13 and 16, contain the supplementary crystallographic data for this paper. These data can be obtained, free of charge, from The Cambridge Crystallographic Data Centre via www.ccdc.cam.ac.uk/structures.
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We thank the EPFL NMR facility staff and especially P. Mieville for help with acquiring 2D spectra, the EPFL XRD facility staff for help with crystal structure determination, and the EPFL MS facility for help with HRMS measurements. We thank O. Naveiras and A. Oggier (ISREC, EPFL), who kindly provided NR-enriched and control diets. We also thank G. Karateev for help and advice regarding the synthesis of TPP-CL2. We appreciate the help of R. Combe and M. Kulagin, members of the CPG facility at EPFL, in measuring the oxygen consumption by mice. We thank the Swiss National Foundation (grant no. 31003A_150134) for funding this work.
The authors declare no competing interests.
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Bazhin, A.A., Sinisi, R., De Marchi, U. et al. A bioluminescent probe for longitudinal monitoring of mitochondrial membrane potential. Nat Chem Biol 16, 1385–1393 (2020). https://doi.org/10.1038/s41589-020-0602-1
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