Abstract
Hydrogen peroxide (H2O2) is emerging as a newly recognized messenger in cellular signal transduction1,2. However, a substantial challenge in elucidating its diverse roles in complex biological environments is the lack of methods for probing this reactive oxygen metabolite in living systems with molecular specificity. Here we report the synthesis and application of Peroxy Green 1 (PG1) and Peroxy Crimson 1 (PC1), two new fluorescent probes that show high selectivity for H2O2 and are capable of visualizing endogenous H2O2 produced in living cells by growth factor stimulation, including the first direct imaging of peroxide produced for brain cell signaling. The combined features of reactive oxygen species selectivity, sensitivity to signaling levels of H2O2, and live-cell compatibility presage many new opportunities for PG1, PC1 and related synthetic reagents for exploring the physiological roles of H2O2 in living systems with molecular imaging.
*Note: In the version of this article initially published, the second author's last name is misspelled. The author's name should read Orapim Tulyathan. The error has been corrected in the HTML and PDF versions of the article.
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Change history
03 May 2007
changed from Tulyanthan to Tulyathan, first n removed
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Acknowledgements
We thank the University of California, Berkeley, the Dreyfus Foundation, the Beckman Foundation, the American Federation for Aging Research, the Packard Foundation, and the US National Institute of General Medical Sciences (NIH GM 79465) for funding this work. E.W.M. thanks the Chemical Biology Graduate Program sponsored by the US National Institutes of Health (T32 GM066698) and a Stauffer fellowship for support. We thank D. Fortin and S. Szobota for providing neuronal cultures, and we thank K. McNeill (University of Minnesota) for the polymer-supported Rose Bengal. Confocal fluorescence images were acquired at the Molecular Imaging Center at UC Berkeley. We also thank A. Fischer at the UC Berkeley Tissue Culture Facility for expert technical assistance.
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E.W.M. performed all of the synthetic and imaging experiments. O.T. prepared the neuronal cultures and helped with the immunostaining experiments. C.J.C. and E.W.M. designed experimental strategies with help from E.Y.I. C.J.C. and E.W.M. wrote the paper.
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Supplementary information
Supplementary Fig. 1
Spectroscopic responses and selectivities for PG1 and PC1 in different buffers. (PDF 107 kb)
Supplementary Fig. 2
Relative responses of PG1, PC1 or DCFH to H2O2 or high-valent iron-oxo centers generated from heme and H2O2. (PDF 239 kb)
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Miller, E., Tulyathan, O., Isacoff, E. et al. Molecular imaging of hydrogen peroxide produced for cell signaling. Nat Chem Biol 3, 263–267 (2007). https://doi.org/10.1038/nchembio871
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DOI: https://doi.org/10.1038/nchembio871
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