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An orange calcium-modulated bioluminescent indicator for non-invasive activity imaging

Nature Chemical Biology volume 15, pages 433–436 (2019)Cite this article

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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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Fig. 1: CaMBI characteristics and imaging of calcium activity with Orange CaMBI reporters.
Fig. 2: Non-invasive calcium imaging in the liver of a live mouse with Orange CaMBI.

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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.).

Author information

Author notes

  1. Younghee Oh

    Present address: Department of DNA analysis, National Forensic Service Seoul Institute, Yangcheon-gu, Seoul, Republic of Korea

  2. Nicole K. Paulk

    Present address: Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA, USA

  3. Namdoo Kim

    Present address: Department of Chemistry, Kongju National University, Gongju, South Korea

Authors and Affiliations

  1. Department of Neurobiology, Stanford University, Stanford, CA, USA

    Younghee Oh, Yunhee Park, Julia H. Cho, Lan Xiang Liu, Namdoo Kim & Michael Z. Lin

  2. Department of Bioengineering, Stanford University, Stanford, CA, USA

    Younghee Oh, Yunhee Park, Julia H. Cho, Lan Xiang Liu, Namdoo Kim & Michael Z. Lin

  3. Department of Pediatrics, Stanford University, Stanford, CA, USA

    Younghee Oh, Yunhee Park, Julia H. Cho, Nicole K. Paulk, Lan Xiang Liu, Namdoo Kim, Mark A. Kay & Michael Z. Lin

  4. Department of Medicine, Stanford University, Stanford, CA, USA

    Haodi Wu & Joseph C. Wu

  5. Department of Radiology, Stanford University, Stanford, CA, USA

    Haodi Wu & Joseph C. Wu

  6. Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA

    Haodi Wu & Joseph C. Wu

  7. Department of Genetics, Stanford University, Stanford, CA, USA

    Nicole K. Paulk & Mark A. Kay

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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.

Corresponding author

Correspondence to Michael Z. Lin.

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The authors declare no competing interests.

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Supplementary information

Supplementary Information

Supplementary Tables 1–4, Supplementary Figures 1–8

Reporting Summary

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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