Abstract
Fluorescent indicators are used widely to visualize calcium dynamics downstream of membrane depolarization or G-protein-coupled receptor activation, but are poorly suited for non-invasive imaging in mammals. Here, we report a bright calcium-modulated bioluminescent indicator named Orange CaMBI (Orange Calcium-modulated Bioluminescent Indicator). Orange CaMBI reports calcium dynamics in single cells and, in the context of a transgenic mouse, reveals calcium oscillations in whole organs in an entirely non-invasive manner.
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Data availability
Nucleotide sequences are available at GenBank for Orange CaMBI 110 (MK558047), Orange CaMBI 300 (MK558048), Blue CaMBI (MK558049), and Green CaMBI (MK558050). Mammalian expression plasmids are available at Addgene for Orange CaMBI 110 (124094), Orange CaMBI 300 (124095), Blue CaMBI (124096), and Green CaMBI (124097). All other data from this study are available from the corresponding author upon request.
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Acknowledgements
We thank members of the Lin laboratory for assistance with experiments, and the Stanford Transgenic, Knockout, and Tumor Model Center for generating the Orange 110 CaMBI transgenic mice. This work was supported by an AHA Postdoctoral Fellowship (to N.K.), an AHA Innovation Grant 15IRG23290018 (to M.Z.L.), a Stanford Discovery Innovation Award (to M.Z.L. and Y.P.), NIH grants R01HL133272 (to J.C.W.), R01HL128170 (to J.C.W.), U01HL099776 (to J.C.W.), and U01NS090600 (to M.Z.L.); NIH fellowship F32HL119059 (to N.K.P.); and NIH Pioneer Award 5DP1GM111003 (to M.Z.L.).
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Authors and Affiliations
Contributions
Y.O. developed CaMBIs, characterized CaMBI properties, performed cellular and animal experiments, performed analysis, and co-wrote the manuscript. Y.P. characterized CaMBI properties, generated trangenic mice, performed cellular and animal experiments, and performed analysis. J.C. and N.K. performed additional characterization of CaMBIs. H.W. assisted in culture of cardiomyocytes. N.K.P. and L.L. assisted with animal experiments. M.A.K. and J.C.W. provided training and advice. M.Z.L. designed experiments, performed analysis, provided training and advice, and co-wrote the manuscript.
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Supplementary information
Supplementary Information
Supplementary Tables 1–4, Supplementary Figures 1–8
Supplementary Video 1
Histamine-induced calcium oscillations in HeLa cells reported by Orange CaMBI 110. Time-lapse images of the cells in Supplementary Fig. 4a are shown.
Supplementary Video 2
Histamine-induced calcium oscillations in HeLa cells reported by Green CaMBI 110. Time-lapse images of the cells in Supplementary Fig. 4b are shown.
Supplementary Video 3
Spontaneous calcium oscillations in a cardiomyocyte reported by Orange CaMBI 110. Time-lapse images of the cells in Fig. 1d are shown.
Supplementary Video 4
Spontaneous calcium oscillations in neurons reported by Orange CaMBI 110. Time-lapse images of the cells in Fig. 1e are shown.
Supplementary Video 5
Spontaneous calcium oscillations in neurons reported by Orange CaMBI 300. Time-lapse images of the cells in Supplementary Fig. 4e are shown.
Supplementary Video 6
Noninvasive imaging of vasopressin-induced calcium oscillations in mouse liver by Orange CaMBI 110 with expression gated by viral-transduced cre. Time-lapse images of the mouse in Fig. 2 are shown.
Supplementary Video 7
Noninvasive imaging of vasopressin-induced calcium oscillations in mouse liver by Orange CaMBI 110 with expression gated by an albumin-cre transgene. Time-lapse images of the mouse in Supplementary Fig. 8 are shown.
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Oh, Y., Park, Y., Cho, J.H. et al. An orange calcium-modulated bioluminescent indicator for non-invasive activity imaging. Nat Chem Biol 15, 433–436 (2019). https://doi.org/10.1038/s41589-019-0256-z
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DOI: https://doi.org/10.1038/s41589-019-0256-z
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